EasyScript cDNA Synthesis Kit

Application:

• First strand cDNA synthesis for PCR
• Construction of cDNA libraries
• Generation of probes for hybridization

EasyScript cDNA Synthesis Kit is a complete system for the efficient synthesis of first strand cDNA from RNA templates. The recombinant RNasin Ribonuclease Inhibitor, supplied with the kit effectively protects RNA template from degradation.

The kit is also supplied with both oligo(dT) and random primers. The oligo(dT) anneals selectively on the poly(A) tail of mRNA. Random primers do not require the presence of poly(A). Therefore, they can be used for transcription of the 5'-end regions of mRNA. Gene-specific primers may also be used with the kit. The first strand of cDNA can be directly used as a template in PCR.

EasyScript Reverse Transcriptase (RTase) within the kit is a genetically modified form of Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV). Both the EasyScript and EasyScript Plus are RNase H deficient (negative). The enzyme is purified from bacteria host as a single holoenzyme of 71 kDa with the capacity of first-strand cDNA synthesis of template up to 9kb and can be used for routine cDNA synthesis.

Kit Components:

Storage Buffer:
50mM Tris-HCl (pH 8.3), 100 mM NaCl, 0.1 mM EDTA, 5 mM DTT, 0.1% (v/v) Triton X-100, and 50% (v/v) glycerol.

Storage: 
Store at -20°C in a frost-free freezer. Multiple freezing and thawing of RNA should be avoided. Keep RNA on ice all the time. It is recommended that the first strand cDNA synthesis is carried out under conditions where RNase contamination has been eliminated.

General Protocol:

RT-PCR reactions should be assembled in a RNA-free environment. The use of "clean", automatic pipettes designated for PCR and aerosol resistant barrier tips are recommended. 
1. Thaw template RNA and all reagents on ice. Mix each solution by vortexing, and centrifuge briefly to collect residual liquid from the sides of the tubes.
2. Prepare the following reaction mixture in a PCR tube on ice:

3. Incubate at 25°C for 10 minutes if random primer is used. Omit this step if Oligo(dT) primer or sequence specific primer are used.
4. Incubate the mixture at 42°C for 60 minutes.
5. Stop the reaction by heating at 85°C for 5 minutes.
6. Chill on ice. The newly synthesized first-strand cDNA now can be used directly for PCR amplification.

Notes:
1. Isolation of poly(A)+ RNA from total RNA is not mandatory; however, doing so may improve the yield and purity of the final product.
2. RNA sample must be free of contaminating genomic DNA.
3. Unlike the oligo(dT) priming, which usually requires no optimization, the ratio of a random primer to RNA is critical in terms of the average length of cDNA synthesized in the reaction. Increasing the ratio of random primer/RNA will result in higher yield of shorter (~500 bp) cDNA, whereas decreasing this ratio will produce longer products.

4. The synthesized cDNA should be stored at -20°C.

EasyScript Reverse Transcriptase

Application:

• Synthesis cDNA froma single-stranded RNA or DNA primer extension
• Sequencing dsDNA
• cDNA library
• Template production for use in PCR
• 3'-end labeling of duplex DNA via end-filling reactions

EasyScript Reverse Transcriptase is a genetically modified form of Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV). Both the EasyScript and EasyScript Plus are RNase H deficient (negative). The enzyme is purified from bacteria h o s t as a single holoenzyme of 71 kDa with the capacity of first-strand cDNA synthesis of template up to 9kb and can be used for routine cDNA synthesis.

Kit Components:

Storage Buffer:
50mM Tris-HCl (pH 8.3), 100 mM NaCl, 0.1 mM EDTA, 5 mM DTT, 0.1% (v/v) Triton X-100, and 50% (v/v) glycerol.

Storage: 
Store at -20°C in a frost-free freezer. Multiple freezing and thawing of RNA should be avoided. Keep RNA on ice all the time. It is recommended that the first strand cDNA synthesis is carried out under conditions where RNase contamination has been eliminated.

General Protocol:

RT-PCR reactions should be assembled in a RNA-free environment. The use of "clean", automatic pipettes designated for PCR and aerosol resistant barrier tips are recommended.

1. Thaw template RNA and all reagents on ice. Mix each solution by vortexing, and centrifuge briefly to collect residual liquid from the sides of the tubes. 
2. Prepare the following reaction mixture in a PCR tube on ice:

3. Incubate at 25°C for 10 minutes if random primer is used. Omit this step if Oligo(dT) primer or sequence specific primer are used.
4. Incubate the mixture at 42°C for 60 minutes.
5. Stop the reaction by heating at 85°C for 5 minutes. 
6. Chill on ice. The newly synthesized first-strand cDNA now can be used directly for PCR amplification.

Notes:
1. Isolation of poly(A)+ RNA from total RNA is not mandatory; however, doing so may improve the yield and purity of the final product. 
2. RNA sample must be free of contaminating genomic DNA.
3. Unlike the oligo(dT) priming, which usually requires no optimization, the ratio of a randomprimer to RNA is critical in terms of the average length of cDNA synthesized in the reaction. Increasing the ratio of random primer/RNA will result in higher yield of shorter (~500 bp) cDNA, whereas decreasing this ratio will produce longer products.

4. The synthesized cDNA should be stored at -20°C.

ENDURO VE20 Vertical Gel Electrophoresis System

Enduro VE20 Vertical Electrophoresis System (20 x 20 cm), includes glass plates, 2 x 24 well combs, cooling coil and casting base

• Large format vertical gel system
• Electroblotting and gel electrophoresis in same universal gel tank
• Quick assembly system for ease of use
• Complete system allows casting of gels and running of gels at same time increasing efficiency

The ENDURO VE20 is Labnet's new large format vertical gel electrophoresis system. It distinguishes itself in its flexibility of applications and ease of gel casting and set-up requiring only 4 screws to secure 4 gels. The same gel tank can house a large format electroblotting assembly sold separately. This system is ideal for first- and second dimension SDS-PAGE, native, preparative, gradient and high-resolution nucleic acid electrophoresis.

Innovative screw-clamp technology consists of only four screws to secure up to four 20 x 20cm gels. This vertical screw clamp technology distributes pressure evenly along the height of gel rather than in the center to eliminate plate bowing and gel compression, while maintaining a leak-proof seal during casting.

NO TANK TOP ASSEMBLY
A built-in inner buffer chamber within the PAGE insert allows set up to be completed without inclusion of an upper buffer chamber.

CASTING ADVANTAGES

• Dual purpose PAGE insert eliminates time consuming transfer of glass plates between separate casting and gel running components.
• Glass plates with bonded spacers assure clean well formation, correct alignment for leak-free casting and eliminates manually aligning spacers.
• Casting stand design ensures leak-free casting even if glass plates are misaligned.

Specifications:

ENDURO™ Semi Dry Blotter

• Western, Southern, and Northern Blots
• Rapid transfer times
• Uniform head dispersion
• Variable gel thickness
• Minimal buffer volume

The ENDURO™ Semi-Dry Blotter provides great flexibility in a lab since it can be used for all types of blotting: Western, Southern, and Northern. Blotting with the ENDURO™ Semi Dry Blotter typically can be done in 15 to 30 minutes making it a fast and easy way to do all your blotting applications. Set up procedures are easy and economical with little buffer needed. The screw down lid adjusts to varying gel thicknesses and sizes while the platinum coated electrodes provide uniform pressure ensuring even transfers. To provide your lab with total blotting solutions use with our ENDURO™ 250V, 3000mA high current power supply.

Specifications

Enduro™ UV Transilluminators

Semi-Dry Blotting Systems, 10cm x 10cm

Accuris MyVolt™ Mini Power Supply

• Compact and simple
• Choose constant 100 V or 200 V output
• Two pairs of output terminals

The Accuris MyVolt Mini is a compact electrophoresis power supply ideal for basic, constant voltage applications. Users can select constant 100 V or 200 V output, and the current can reach a maximum of 200 mA.

Two sets of output jacks are available to connect up to 2 gel rigs and these small power supplies can securely be stacked for use with additional gel rigs.

Specifications

Wide Format Gel System 23.5 cm x 14 cm

•     Heavy duty lower buffer chamber
•     Interlock safety lid with attached leads
•     Non-gasketed 23cm wide x 14cm long UV-transparent gel tray with built-in fluorescent ruler and 4 comb slots.
•     Four 1.5mm x 50 tooth combs
•     Opaque well visualization plate.